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Cho, Yeo, You, Chang, Moon, and Kim: The Expression of p53 and Phosphorylation of H2AX in Germ Cells of Varicocele Rats
Original Article
Korean Journal of Urology

Korean Journal of Urology 2006; 47(12): 1354-1360.

Published online: 31 December 2006

DOI: https://doi.org/10.4111/kju.2006.47.12.1354

The Expression of p53 and Phosphorylation of H2AX in Germ Cells of Varicocele Rats

Won Jin Cho, Moon Hwan Yeo1, Ho Jin You1, 2, In Youb Chang1, 3, Hyung Yoon Moon, Chul Sung Kim

Department of Urology, College of Medicine, Chosun University, Gwangju, Korea.

1Research Center for Proteinous Materials, College of Medicine, Chosun University, Gwangju, Korea.

2Department of Pharmacology, College of Medicine, Chosun University, Gwangju, Korea.

3Department of Anatomy, College of Medicine, Chosun University, Gwangju, Korea.

Corresponding author (mu-hn@hanmail.net)

Received 10 July 2006       Accepted 29 August 2006

Copyright © 2006 The Korean Urological Association

Abstract

Purpose

To explore the expressions of p53 and phosphorylation-H2AX in varicocele-induced rat testes.

Materials and Methods

A total of 16 adult male Sprague-Dawley rats underwent an operation; 12 underwent an experimental varicocele and 4, as controls, were sham-operated. Groups of 4 varicocele-induced rats underwent a left orchiectomy after 2 or 3 weeks, or both orchiectomies after 4 weeks. The sham-operated rats underwent both orchiectomies after 4 weeks. Sections of both testes from each animal were studied. The changes in the expressions of p53 and phosphorylation of H2AX were determined using immunohistochemistry and western blot.

Results

Immunohistochemical staining of the left testes in the varicocele-induced rats showed that the expressions of p53 and phosphorylation of H2AX had not begun 2 weeks postoperatively, but remarkable results were observed after 3 and 4 weeks. Both testes of the varicocele-induced rats showed the expressions of p53 and phosphorylation of H2AX after 4 weeks, with the left testes being more distinctive in immunohistochemical staining compared to the right. Western blot of the left testes in the varicocele-induced rats also showed unclear expressions of p53 and γ-H2AX after 2 weeks. Considerable distinction was seen after 3 and 4 weeks compared to the control group.

Conclusions

Our results suggest that experimental varicocele is associated with increased sperm DNA damage. These changes may be related to abnormal spermatogenesis.

Keywords: DNA damage, Varicocele, p53, γ-H2AX

Figures and Tables

Fig. 1
Immunohistochemical staining for p53 in varicocele-induced rats. Expressions of p53 at 3 and 4 weeks (B, C and D) are increased compared to the control group (E and F). (A) Left testis of the varicocele-induced rat after 2 weeks, (B) Left testis of the varicocele-induced rats after 3 weeks, (C) Left testis of the varicocele-induced rats after 4 weeks, (D) Right testis of the varicocele-induced rats after 4 weeks, (E) Left testis of the sham-operated rats after 4 weeks (control group), (F) Right testis of the sham-operated rats after 4 weeks (control group).
kju-47-1354-g001
Fig. 2
Immunohistochemical staining for γ-H2AX in the varicocele rats. Expressions of phosphorylation of H2AX after 3 and 4 weeks (B, C and D) are increased compared to control group (E and F). (A) Left testis of the varicocele-induced rat after 2 weeks, (B) Left testis of the varicocele-induced rats after 3 weeks, (C) Left testis of the varicocele-induced rats after 4 weeks, (D) Right testis of the varicocele-induced rats after 4 weeks, (E) Left testis of the sham-operated rats after 4 weeks (control group), (F) Right testis of the sham-operated rats after 4 weeks (control group).
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Fig. 3
Western blotting of the p53 and γ-H2AX expressed in the varicocele-induced rat testes. Protein extracted from the rat testis, with (2, 3 and 4 weeks) or without (4 weeks) partial ligation of the Lt renal vein, is analyzed for the expression levels of p53, γ-H2AX and actin by Western blotting. The expressions of p53 and γ-H2AX after weeks 3 and 4 are significantly higher compared to control group and the varicocele-induced rats 2 weeks postoperatively.
kju-47-1354-g003

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